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1.
AIM: To investigate the effect of Sonic Hedgehog (Shh) signaling blockade on the growth of hematocarcinoma cells and underlying mechanisms. METHODS: The expression of Shh signaling molecules in hematocarcinoma cell lines BEL-7402, Huh7 and HepG2 was detected by RT-PCR. The cell viability was detected by MTT assay. The cell cycle and apoptosis were analyzed by flow cytometry. The expression of apoptosis-related proteins was determined by Western blot. RESULTS: Shh signaling molecules were all expressed in BEL-7402, Huh7 and HepG2 cells. The mRNA expression of Patched (Ptch), Gli1 and Gli2 was down-regulated by anti-Shh antibody. Blockade of Shh signaling pathway inhibited the proliferation of hepatocarcinoma cells with increasing cells in G0/G1 phase and induced the apoptosis of hepatocarcinoma cells. Treatment with anti-Shh antibody down-regulated the protein expression of pro-caspase-3, pro-caspase-8 and pro-caspase-9, while up-regulated the protein levels of cleaved caspase-3, cleaved caspase-8 and cleaved caspase-9 in BEL-7402 cells. CONCLUSION: Blockade of Shh signaling pathway inhibits the growth of hepatocarcinoma at different levels by cell cycle arrest and inducing apoptosis of hematocarcinoma cells.  相似文献   

2.
AIM: To investigate the produce of intracellular cytokine following short-term in vitro stimulation with vMIP and LPS, and discuss the effect of vMIP to cellular immunity. METHODS: The methods of Cross-linking of radioactivity, ELISA and four-colors flow cytometer were used to test the level of the secretion of chemokine IL-12 and intracellular cytokine IFN-γ and IL-4. RESULTS: After treated the PBMCs with vMIP-II, the levels of secretion of IL-12, IFN-γ and IL-4 were reduced in the present of LPS by competitively combining chemokine receptor; vMIP promoted CD4+T cell to secrete IL-12, IFN-γ and IL-4. CONCLUSION: vMIP-II can protect systemic response of immunity and reduce extremely inflammation by down-regulating proinflammation.  相似文献   

3.
AIM: To explore the effects of sterigmatocystin (ST) on IL-2 and IFN-γ expression and secretion in murine spleen cells in vitro. METHODS: The secretion and expression of IL-2 and IFN-γ in murine spleen cells after ST pretreatment at five different dosages(0.125 mg/L,0.25 mg/L,0.5mg/L,1 mg/L,2 mg/L) were studied with ELISA and semi-quantitative RT-PCR method, respectively. RESULTS: Pretreatment of murine spleen cells in vitro with ST at five different dosages affected the IL-2 and IFN-γ secretion at protein level and expression at mRNA level of the treated cells. The effects varied dependently to the ST dosage. At relatively lower dosages, ST induced the expression of IL-2 and IFN-γ in murine spleen cells, while at relatively higher dosages, inhibitory effects were found, with the most significant inhibitory effects seen in ST 1 mg/L group. CONCLUSION:ST affected the se-cretion and expression of IL-2 and IFN-γ in treated murine spleen cells.At relatively lower dosage, ST induced IL-2 and IFN-γ secretion and expression, while at relatively higher dosages，inhibitory effects appeared.  相似文献   

4.
AIM:To investigate the influences of protein kinase C(PKC) inhibitors on the expression of interleukin-2(IL-2) and interferon-γ(IFN-γ) byin vitro activated T-lymphocytes. METHODS:Double fluorescent staining together with flow cytometry was adopted to detect intracellular cytokines and to analyze the effects of H7 and gossypol on IL-2 and IFN-γ expression levels of T-lymphocytes stimulated with phorbol ester (PDB)+ionomycin(I) in the presence of monensin.RESULTS:The expression rates of IL-2 and IFN-γ of CD3+ T cells stimulated with PDB+I for 4 h were 16.64±2.04 and 25.81±3.53(x±s), respectively, which were significantly higher than that of control (1.06±0.22 and 3.12±0.77)(P＜0.05). Gossypol was able to inhibit the expression of IL-2 and IFN-γ significantly, with the expression rates of 2.08±0.12 and 9.01±1.90, respectively. At the presence of 50 μmol/L H7, the rates of IL-2+ and IFN-γ+ CD3+ T cells were 0.43±0.06 and 2.40±0.27, respectively. The effect of H7 was stronger than that of gossypol. CONCLUSION:PKC plays an important role in the expression of IL-2 and IFN-γ of CD3+T cells and its inhibitors H7 and gossypol exert significant inhibitory effect on the expression of these two cytokines. It is suggested that H7 and gossypol may have modulatory effect on T-cell-dependent specific immune responses by inhibiting PKC activity.  相似文献   

5.
AIM: To investigate the effect of R848 (a Toll-like receptor 7/8 agonist) combined with poly-inosinic:polycytidylic acid[Poly(I:C), a Toll-like receptor 3 agonist] on dendritic cell (DC) maturation, and the killing effect of DC-induced cytotoxic T-lymphocytes (CTL) on human lung adenocarcinoma A549 cells. METHODS: Mononuclear cells were isolated from human peripheral blood and induced to differentiate into DC. The whole-cell lysate of A549 cells, namely tumor cell lysate (TCL), was used as antigen. R848 combined with Poly(I:C) was used as adjuvant to stimulate the DC. DC surface markers were analyzed by flow cytometry. The DC stimulated by antigen was co-cultured with T-lymphocytes for 7 d to induce CTL. The culture supernatant and CTL were collected. The levels of interleukin-12 (IL-12) p70, interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in the supernatant were measured by ELISA. The CTL and A549 cells were co-cultured for 16 h, and the cytotoxicity was observed by LDH assay.RESULTS: The expression of CD83 and CD80 on the DC surface, and the secretion of IL-12 p70 in DC-R848+Poly(I:C) group were significantly increased compared with DC-TCL group (P<0.01). In addition, the cytotoxicity of CTL for A549 cells in DC-R848+Poly(I:C) group was significantly enhanced compared with DC-TCL group (P<0.01). The secretion levels of IFN-γ and TNF-α in DC-R848+Poly(I:C) group were significantly elevated compared with DC-TCL group (P<0.01). CONCLUSION: R848 combined with Poly(I:C) significantly promotes DC maturation and activation, and enhances the antigen-presenting effect of DC and the cytotoxicity of DC-induced CTL.  相似文献   

6.
AIM: To investigate the effect and mechanism of Heligmosomoides polygyrus (H. polygyrus) infection in mouse inflammatory bowel disease (IBD) mediated by CD4+ helper T-cells. METHODS: Ovalbumin (OVA) -specific CD4+ helper T-cells were transferred into SCID (severe combined immunodeficiency) mice to establish an IBD model. The IBD mice were infected by H. polygyrus and sacrificed 14 days later. The histological changes of the colon were observed, and the expression of interferon-gamma (IFN-γ) and interleukin-4 (IL-4) in mesenteric lymph nodes was detected by ELISA and flow cytometry. Additionally, IL-4 monoclonal antibody was intraperitoneally injected into the H. polygyrus-infected IBD mice to block the secretion of IL-4. The IL-4-blocking IBD mice were sacrificed 9 days later and the above indexes were also determined.RESULTS: Compared with the non-infection group, the H. polygyrus-infected IBD mice had more severe colonic lesions, higher level of IL-4 and lower level of IFN-γ in mesenteric lymph nodes (all P<0.05). Compared with the non-blocking group, the H. polygyrus-infected IBD mice with IL-4 blockage had less colonic lesions, lower IL-4 level and higher IFN-γ level (all P<0.05).CONCLUSION: H. polygyrus infection in CD4+ T-cell-mediated IBD model promotes inflammation in the early stage probably by inducing the secretion of Th2 cytokine and inhibiting the secretion of Th1 cytokine. The finding suggests that using worms for treatment of IBD needs to be cautious.  相似文献   

7.
AIM: To investigate the effect and mechanism of Foxp3-transduced CD4+CD25-T cells on the cytotoxicity of NK cells. METHODS: Retroviral Foxp3 gene transfection was applied to nave CD4+CD25-T cells. Fresh transduced CD4+Foxp3+ T cells were co-cultured with NK cells. ［51Cr］ labeled YAC-1 cells were used to detect NK cells cytotoxicity. The anti-TGF-β antibody was added into the co-culture system to detect the TGF-β blocking effect. Also the transwell co-culture system was used to investigate the regulatory effect of Treg cells on NK cells. RESULTS: One week after transduction, 38.0% of Foxp3-transduced T cells showed GFP expression by flow cytometry. Foxp3-transduced CD4+CD25-T cells suppressed function of NK cells. The inhibition rates of Foxp3 transduced CD4+CD25- T cells were 42.9% at 24 h and 22.7% at 48 h. When anti-TGF-β antibody was added to the co-culture system, the inhibition rate of CD4+Foxp3+ T cells was 3.2% and 2.1%, respectively. CONCLUSION: CD4+Foxp3+ T cells significantly inhibit the cytolytic function of NK cells. TGF-β plays different roles on this action in different inhibition systems. The inhibitory effect of Treg cells on NK cells is cell-to-cell contact dependent and associates with TGF-β expression.  相似文献   

8.
AIM: To observe the effect of B7H1 expression in pancreatic carcinoma cells on the proliferation and activation of co-cultured T lymphocytes. METHODS: B7H1 expression in panc-1 cells before and after interferon-γ(IFN-γ) treatment or B7H1-siRNA transfection was evaluated by RT-PCR and flow cytometry. The influence of B7H1 expression on co-cultured PHA-activated T lymphocytes was determined by the methods of MTT and enzyme-linked immunosorbent assay (ELISA). RESULTS: B7H1 was highly expressed in panc-1 cells and up-regulated after IFN-γ stimulation. Such up-regulation led to the significant inhibition of T cell proliferation and secretion of cytokines such as IFN-γ and interleukin-2(IL-2). However, IL-10 production was enhanced. In contrast, knockdown of B7H1 expression in panc-1 cells by RNA interference resulted in increased T cell proliferation as well as IFN-γ and IL-2 production. Meanwhile, the IL-10 secretion decreased. CONCLUSION: B7H1-expressing panc-1 cells suppress T cell function by inhibiting T cell proliferation and production of Th1 cytokines. Suppression of B7H1 expression through siRNA restores T cell immune functions, indicating a potential strategy for immunotherapy against pancreatic cancer.  相似文献   

9.
AIM:To study the effect of IFN-γ inhalation on the anti-infection ability of the lungs in the immunocompromised host. METHODS:The immunological factors in the immunocompromised rats and the immunocompromised rats administrated IFN-γ via aerosol were investigated after 1, 3, 7 days when they were injected Candida albicans via tracheal. The Canidda albicans count of the left lung was also determined after 7 days when injecting pathogen. RESULTS:The Canidda albicans count of the left lung in IFN-γ group was significantly less than that of control group. The phagocyting and bactericidal percentages, Ia antigen expression percentages, the levels of TNF-α, IL-1β and IL-6 in the culture supernatant of the AM, the activity of IFN-γ and TNF-α in BALF (except the TNF-α on 7 th day) in IFN-γ group were markedly higher than those in control group. The expression of IFN-γ and IL-1β pulmonary tissues in IFN-γ group was higher than that in control group. The expression of TNF-α in IFN-γ group was less than that in control group. The expression of IL-6 was no changes between two groups. The levels of IFN-γ, IL-1β and IL-6 in the blood (except IL-1β on 3 rd day), and the killing ability of the lymphocytes in blood had no difference between two groups. CONCLUSION:Administration of IFN-γ via aerosol obviously enhanced the anti-infection ability of the lungs in the immunocompromised host, but has no influence on the whole body cellular immunity.  相似文献   

10.
AIM: To investigate the molecular mechanism and the immunosuppressive phenotype of macrophages under long-term exposure to lipopolysaccharide (LPS). METHODS: We used Ficoll-Hypaque density gradient centri-fugation combined with MicroBeads Separation Kits to separate peripheral blood mononuclear cells from human blood, and then induced the monocytes into macrophages. We observed the morphology of the macrophages by treating the cells with LPS for 48 h, in comparison with a negative control and IFN-γ treatment. ELISA was used to detect the levels of cytokines, such as IL-10, IL-12, IL-6 and TNF-α, and flow cytometry was used to detect the expression of the surface molecules (HLA-DR, CD14, CCR7, HLA-ABC and CD40). To observe the effect of macrophage on T cell proliferation, co-culture experiment was carried out for 6 d. Real-time PCR was used to validate the expression levels of molecules related to MyD88-independent pathway in Toll-like receptor 4 (TLR4) signal pathway. RESULTS: The antigen-presenting ability of the macrophages was reduced and the IL-10 expression level was increased after the cells were treated with LPS for 48 h. We observed a poor proliferative capacity of CD8+ T cells after co-culturing of LPS-induced macrophages with CD3+ T cells for 6 d. The results of real-time PCR indicated that TRIF, IRF3 and CIITA were down-regulated in LPS-induced macrophages.CONCLUSION: We successfully established a macrophage model in vitro and observed that LPS-induced macrophages into an immunosuppressive phenotype with poor CD8+ T cell proliferative capacity, in which MyD88-independent TLR4 signaling pathway was impaired.  相似文献   

11.
AIM: To investigate the amount and patterns of expressing CD69, IL-4 and IFN-γ on TCRVα24+ NKT cells, and compare with that of CD3+ T cells from human peripheral blood in response to in vitro stimulation. METHODS: The whole blood was stained with three-color immunofluorescence directly or after cultured with PDB+ionomycin (Ion) for 6 h, then the mononuclear cells were separated by lysing red blood cells. The expression rates of CD69, IL-4 and IFN-γ on TCRVα24+ NKT cells and CD3+ T cells were estimated by flow cytometer. RESULTS: As a proportion of mature T cells, the ratio of TCRVα24+ NKT cells to CD3+ T cells was about (1.34±0.42)%. The expression rates of CD69 on TCRVα24+ NKT cells and CD3+T cells in response to PDB+Ion for 6 h were (96.71±1.33)% and (98.60±0.47)%, respectively, while the ratio were (11.47±2.86)% and (1.07±0.45)% in the unstimulated group, and there were significant difference between them. The expression rates of IL-4 and IFN-γ on TCRVα24+ NKT cells stimulated with PDB+Ion for 6 h were (48.62±2.44)% and (46.65±8.91)%, respectively, which were significantly higher than that of unstimulated group [(31.57±3.31)%, (13.45±6.29)%] and that of stimulated CD3+ T cells, though the expression rates on stimulated CD3+ T cells were significantly higher than that of unstimulated CD3+ T cells. CONCLUSION: There is small amount of NKT cells in adult human peripheral blood. The expression rates of IFN-γ and IL-4 on these lymphocytes are higher than CD3+ T cells, suggesting that NKT cells are important immunomodulatory cells in special microvironments.  相似文献   

12.
AIM: To investigate the effect of mesalazine treatment on regulation of Th1, Th17 and Treg cells in mice with dextran sulfate sodium (DSS)-induced ulcerative colitis (UC). METHODS: The expression of IL-17, IFN-gamma and Foxp3 in the peripheral blood mononuclear cells (PBMC) and intestinal mucosa lamina propria mononuclear cells (LPMC) of DSS-induced UC mice was detected by flow cytometry analysis. The effect of mesalazine treatment on regulaiton of Th1, Th17 and Treg cells in the mice with DSS-induced ulcerative colitis was examined.RESULTS: The expression of IL-17, IFN-γ and Foxp3 on CD4+T cells were significantly higher in the PBMC of DSS-induced mice than those in control group. CD4+ IFN-γ+T cells and CD4+ Foxp3+T cells were higher in LPMC than those in control group, except CD4+IL-17+T cells. Moreover, the Th1, Th17 and Treg cells were higher in DSS group than those in control group in LPMC. However, only Tregs was higher in PBMC. Pre-treatment with mesalazine significantly decreased the number of Th17, Th1 and Treg cells of UC model mice both in PBMC and LPMC.CONCLUSION: The Th1, Th17 and Tregs cells in DSS-induced mice were significantly higher than those in control mice, suggesting that CD4+T cell subsets play an important role in the pathogenesis of UC. Mesalazine may play a role in the treatment of UC by regulating the Th1, Th17 and Tregs cells.  相似文献   

13.
AIM: To observe the immunoregulatory effects of prostaglandin E2 receptor (EP) subtypes EP2/EP4 on the B-cells of collagen-induced arthritic(CIA)mice. METHODS: DBA/1 mice were immunized with chicken type II collagen emulsified in Freunds complete adjuvant to induce arthritis. B-cells were isolated from the splenocyte suspension by positive selection using anti-CD19 monoclonal antibody immunomagnetic beads. The expression of MHC II, CD 80 and CD86 was examined by flow cytometry. The mRNA levels of EPs, interferon γ (IFN-γ), tumor necrosis factor α (TNF-α), IL-6, IL-4, IL-10 and transforming growth factor-β (TGF-β) were detected by real-time RT-PCR. RESULTS: The rank of the mRNA levels of EPs was EP2>EP1>EP3>EP4 in B-cells and EP2/EP4 mRNA expression was obviously increased in CIA mice. EP2 antagonists inhibited the expression of MHC II, CD80 and CD86. EP4 antagonist had little effect on CD80. EP2/EP4 antagonists inhibited the mRNA expression of IFN-γ, TNF-α, and IL-6 (P<0.05 or P<0.01) and increased the expression of IL-10 (P<0.01 or P<0.05). Furthermore, the antagonists of EP2 and EP4 also increased the mRNA expression of IL-4 and TGF-β (P<0.01), respectively. CONCLUSION: PGE2 modulate the pathogenesis of CIA via EP2/EP4 by regulating the expression of surface molecules and cytokines in B-cells. EP2/EP4 may be a new therapeutic target for treating rheumatoid arthritis.  相似文献   

14.
AIM: To investigate the influence on the killing effect of NK cells in vitro by up-regulation of human leukocyte antigen-E (HLA-E) expression in hepatocellular carcinoma Bel7402 cells. METHODS: The recombinant lentiviral vector (Lentivirus/CMV/GFP-HLA-E) was constructed and transfected into hepatocellular carcinoma Bel7402 cells. The HLA-E gene expression at mRNA and protein level was monitored by the methods of real-time RT-PCR and Western blotting. The influence on the killing effect of NK cells in vitro by up-regulation of HLA-E expression in hepatocellular carcinoma Bel7402 cells and by HLA-ABC antibody blocking the site on the surface of target cells was analyzed.RESULTS: Real-time RT-PCR showed that there was a significant increase in HLA-E mRNA level in hepatocellular carcinoma Bel7402 cells transfected with Lentivirus/CMV/GFP-HLA-E at 24 h (P<0.05) and 48 h,72 h, 96 h (P<0.01) as compared to blank group. There was also a significant increase in exogenous/endogenous HLA-E proteins at 12 h, 24 h, 48 h, 72 h and 96 h by Western blotting (P<0.01). Without HLA-ABC antibody blocking, there was a statistical difference for the killing effect of NK cells,comparing Bel7402 Lenti HLA-E group with Bel7402 group (P<0.05). Comparing HLA-ABC antibody blocking group with no HLA-ABC antibody blocking group, a statistical difference for the killing effect of NK cells (P<0.05) was observed. There was also a statistical difference for the killing effect of NK cells between Bel7402 with blocking group and Bel7402 Lenti HLA-E with blocking group (P<0.05). CONCLUSION: The vector of Lentivirus/CMV/GFP-HLA-E has an active up-regulation effect in HLA-E mRNA level and HLA-E protein level. While up-regulation of HLA-E in target cells, the killing effect of NK cells on target cells is obviously weakened. Blockage of the sites on the surface of target cells by HLA-ABC antibody universally enhances the killing effect of NK cells on the target cells.  相似文献   

15.
无 《园艺学报》2012,39(1):142-146
自2008年11月在中国农业科学院郑州果树研究所成功召开园艺植物染色体倍性操作与遗传改良研讨会以来,我国科研人员在该领域的研究取得了可喜的成绩。为进一步总结近几年来在该领域的研究进展,促进园艺植物倍性育种研究,同时为该领域的专家、学者和同仁们提供良好的交流平台。中国园艺学会定于2012年4月中旬在重庆召开园艺植物染色体倍性操作与遗传改良学术研讨会。欢迎从事该研究领域及相关研究工作的人员参加。  相似文献   

16.
AIM: Detection and enrichment of T lymphocytes after allogeneic PBMNC stimulation according to secreted cytokine were performed in order to explore a new approach for studying allogeneic reactive T lymphocytes. METHODS: The novel cytokine secretion assay (CKSA) was applied to detect T lymphocyte secreting IFN-γ, IL-4 and IL-10 at single cell level in human mixed lymphocyte reaction. IFN-γ secreting T cells were enriched by means of magnetic sorting system. RESULTS: Allogeneic PBMNC stimulation didn't alter the proportion of IL-4 and IL-10 secreting T lymphocytes (which were 0.12%±0.03% and 0.10%±0.03%, respectively), but increased proportion of IFN-γ secreting T lymphocytes (1.12%±0.13%). These IFN-γ- secreting T lymphocytes could be further enriched to 67.3%±10.5% . CONCLUSION: It is feasible to detect significantly increased IFN-γ-secreting T cells after allogeneic PBMNC stimulation based on the novel CKSA technique, and these cells could be efficiently enriched for further use.  相似文献   

17.
AIM:To investigate the expression of Th1-typed cytokine IFN-γ and Th2-typed cytokine IL-4 on T lymphocytes that infiltrate in nasal polyps for searching the pathogenesis of nasal polyps. METHODS:Nasal polyps tissue samples and peripheral blood were obtained from 21 patients. Normal human inferior turbinate mucosa and peripheral blood were obtained as well. Flow cytometry was adopted to detect the expression of IFN-γ and IL-4 of T lymphocytes. RESULTS: Th cytokines were rarely detected in inferior turbinate from normal human. Nasal polyps tissue consisted of abundant T lymphocytes. The expression of IL-4 and IFN-γ increased in peripheral blood from patients compared with normal human (P<0.05). The expression of IL-4 increased but the expression of IFN-γ decreased in nasal polyps compared with that of peripheral blood from the same patient (P＜0.05). CONCLUSION:There were generous of T lymphocytes infiltrating in nasal polyps. There was abnormal immune status in the local nasal mucosa from the patients, and the predomination of Th cytokine secretion changed compared with peripheral blood from the same patients, which resulted in the change of microenvironment of nasal mucosa and possibly close related to the formation of nasal polyps.  相似文献   

18.
AIM: To detect interleukin 15 (IL-15) levels in peripheral blood from patients with active lupus nephritis and investigate its clinical significance. METHODS: IL-15 level was determined by enzyme linked immunosorbent assay (ELISA). The peripheral blood mononuclear cells (PBMCs) were isolated with grads density abaxiality. The inhibitory effects of dexamethasone on production of IL-15, IgG and anti-dsDNA antibody in cultured PBMCs from LN patients were also investigated. RESULTS: (1) Serum IL-15 level in LN patients was significantly higher than that in normal controls (P<0.01). Serum IL-15 level in active LN patients was significantly higher than that in remised patients (P<0.05). (2) Serum IL-15 level was positively correlated with SLEDAI, anti-dsDNA antibody and 24 h urine protein excretion in active LN patients. (3) Serum IL-15 level was significantly reduced in LN patients treated with combination of cyclophosphamide (CTX) and steroid for 12 weeks. (4) Secretion of IL-15, IgG and dsDNA antibody in cultured PBMCs from active LN patients was significantly higher than that in normal control group, and IL-15 level in supernatant of cultured PBMCs from active LN patients was positively correlated with IgG and dsDNA antibody. Dexamethasone inhibited the secretion of IL-15, IgG and dsDNA antibody in cultured PBMCs from LN patients. CONCLUSION: Serum IL-15 level in patients with active lupus nephritis is significantly elevated, suggesting that IL-15 may be involved in the pathophysiological process of LN. IL-15 may be used as an index to assess the activity of LN.  相似文献   

19.
AIM: To study the effect of hepatitis virus B proteins on peripheral blood mononuclear cells (PBMCs) from patients among various types of chronic hepatitis B virus (HBV) infection.METHODS: 80 patients of various types of chronic HBV infection were observed, including 40 HBeAg positive with abnormal alanine aminotransferase (ALT) (A group), 20 HBeAg positive with persistent normal ALT(B group), 20 HBeAg and HBV-DNA negative with persistent normal ALT level(C group). IL-10, IFN-γ in CD8+CD28+T cells, after stimulation with PHA, HBeAg and HBcAg for 48 h, were inspected respectively in PBMCs.RESULTS: IFN-γ was significantly lower in HBeAg positive patients. IL-10 was significantly higher in HBeAg positive with normal ALT. CD8+CD28+T were significantly lower than others. CONCLUSION: In HBeAg positive group, secretion of cytotoxic T lymphocyte (CTL) and Th1 type cellular immunologic reaction is decreased, Th2 type cellular immunologic reaction is enhanced.  相似文献   

20.
AIM: To investigate how human adipose-derived stem cells (hASCs) regulates the differentiation of Th17 cells in multiple sclerosis. METHODS: hASCs were isolated from the adipose tissues. Magnetic-activated cell sorting (MACS) kit was used to isolate CD4+ T cells from peripheral blood mononuclear cells (PBMCs) which were isolated by density gradient centrifugation. The percentage of CD4+ T cells was detected by flow cytometry. The activated CD4+ T cells were co-cultured with hASCs for about 4 d at different ratios of hASCs to CD4+ T cells (1:4 and 1:10) in a Th17 polarised condition. Another group adding anti-leukemia inhibitory factor (LIF) antibody was set up. Th17 cell proportion of the CD4+ T cells was determined by flow cytometry. The level of LIF in the supernatant of co-cultured system was measured by ELISA. The mRNA expression of retinoid-related orphan receptor γt (RORγt), interleukin-6 receptor (IL-6R), interleukin-23 receptor (IL-23R), LIF and leukemia inhibitory factor receptor (LIFR) was detected by real-time PCR. RESULTS: The result of flow cytometry suggested there were mainly hASCs, and the percentage of CD4+ T cells in the PBMCs were above 90% after MACS. The Th17 cell proportion decreased in 1:4 and 1:10 co-cultured groups in a dose-dependent manner. The mRNA expression of IL-6R, IL-23R and RORγt was downregulated and the expression of LIFR and LIF was up-regulated. When the anti-LIF was added into the co-cultured system, the ratio of Th17 cells increased and reached to the control level. The protein level of LIF obviously increased after co-cultured. After anti-LIF added, the mRNA expression of RORγt and IL-6R was up-regulated. CONCLUSION: hASCs inhibits the differentiation of Th17 cells from multiple sclerosis patients through the competitive inhibition of LIF/IL-6 by secreting LIF.  相似文献   

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